68 research outputs found
A rapid method of Sertoli cell isolation by DSA lectin, allowing mitotic analyses
No full textWe have developed a rapid and convenient method of Sertoli cell preparation for studying the growth kinetics of these cells in in vitro culture. Datura Stramonium agglutinin (DSA)-coated dishes were used to rapidly purify single Sertoli cells from immature rat testis. We have monitored by immunohistochemical markers the degree of contamination of our Sertoli cell preparation by other cell types. The cell preparation is essentially free of germ cells and interstitial cells and contains a minimal percentage of myoid cells. Sertoli cells isolated with this method retain functional activities such as the FSH responsiveness in terms of cAMP production. In addition, we have studied the proliferative activity of Sertoli cells isolated by lectin binding from rats of different ages. Sertoli cells exhibited a characteristic pattern of proliferation which was a function of the donor animal age. The proliferative activity of isolated Sertoli cells decreased with age, being much higher in 3 day-old rats than in older animals. A similar pattern was observed when the mitotic activity of Sertoli cells in response to mitogens present in the testicular extracts from 5 day-old rats was evaluated. The method described here reduces or eliminates many of the drawbacks of the conventional procedures used to isolate Sertoli cells, thus providing a useful tool in studies of growth kinetics and regulation of cell proliferation in vitro. (C) 1998 Elsevier Science Ireland Ltd. All rights reserved
Steroidogenic potential of human fetal kidney at early gestational age: Steroidogenic potential of human fetal kidney at early gestational age
No full textISI Document Delivery No.: IM1YR Times Cited: 0 Cited Reference Count: 25 Savchuk, I. Morvan, M. L. Antignac, J. P. Kurek, M. Le Bizec, B. Soder, O. Svechnikov, K. Frimurare Barnhuset Foundation; Kronprinsessan Lovisas Foundation; Sallskapet Barnavard; Stiftelsen Sigurd och Elsa Goljes Minne; Stiftelsen Olle Engkvist ByggmastareSwedish Research Council; Stiftelsen Gunvor och Josef Aners; Stiftelsen Jane och Dan Olssons; Stiftelsen Tornspiran This work supported by the Frimurare Barnhuset Foundation, Kronprinsessan Lovisas Foundation, the "Sallskapet Barnavard"; the "Stiftelsen Sigurd och Elsa Goljes Minne", the "Stiftelsen Olle Engkvist Byggmastare", the "Stiftelsen Gunvor och Josef Aners", the "Stiftelsen Jane och Dan Olssons" and the "Stiftelsen Tornspiran". 0 3 Elsevier science inc New york 1878-5867International audienceSteroidogenic potential of the human fetal kidney (hFK) at the end of first trimester is poorly investigated. Little is known about the ontogeny of steroidogenic enzymes and activities of steroidogenic pathways in the hFK at early pregnancy. Our aim was to explore steroidogenesis and the expression of steroidogenic enzymes in the hFK at gestational weeks (GW) 9-12. Steroids in the hFK were analyzed by gas chromatography/coupled to tandem mass spectrometry. The expression of steroidogenic enzymes in the hFK at GW 9-12 was investigated by qPCR, automated Western blotting and immunohistochemistry. We observed that the hFK produced substantial amount of steroids of the Delta(5) and Delta(4) pathways and several steroid precursors in the biosynthesis of DHT via the backdoor pathway but not DHT itself. The levels of steroids and expression of relevant steroidogenic enzymes (e.g., CYP17A1, HSD3B1, HSD3B2, CYP11B1 and AKR1C4) we significantly higher in the hFK at GW11-12 compared to GW9. We also found the expression of sex steroid receptors (e.g., AR, ER alpha and ER beta) in the hFK at GW9-12. No sex dependent differences in the levels of all identified steroids and expression of steroidogenic enzymes in the hFK from male and female fetuses were found. Altogether, our data indicate that the hFK at early pregnancy is steroidogenic organ with potential to synthesize multiple steroids that may play an important role in the formation and development of this organ in humans
Interleukin-18 is expressed in rat testis and may promote germ cell growth
No full textMona-Lisa Strand, Aida Wahlgren, Konstantin Svechnikov, Cecilia Zetterstroma, Brian P. Setchell, Olle Sode
Endocrine Disruptors: A Most Relevant Issue for the Pediatric Endocrinologist.
No full textpeer reviewe
Single subcutaneous administration of chorionic gonadotropin to rats induces a rapid and transient increase in testicular expression of pro-inflammatory cytokines
No full textMonique Assmus, Konstantin Svechnikov, Mikael Von Euler, Brian Setchell, Taranum Sultana, Cecilia Zetterström, Mikael Holst, Wieland Kiess and Olle Söde
Role of testicular interleukin-1alpha tIL-1alpha in testicular physiology and disease
No full textThis review focuses on the role of the cytokine interleukin-1alpha (IL-1alpha) in the testis; elaborating upon its importance during the complex process of spermatogenesis while relating this cytokine to some of the pathophysiological states affecting the testis. IL-1alpha, a proinflammatory cytokine, is expressed constitutively by the intact adult rat testis where it acts on germ, Sertoli and Leydig cells to regulate germ cell proliferation and steroidogenesis. The sequence identity of testicular IL-1alpha matches with the one secreted by activated macrophages in systemic immunity. The classical macrophage IL-1alpha is produced as 32 kDa precursor protein which is processed to mature 17 kDa IL-1alpha and a 16 kDa propiece. The rat testicular IL-1alpha, mainly secreted by Sertoli cells, was found to have molecular heterogeneity that can be observed both at the transcriptional and the translational levels. In the rat testis, two transcripts were found to be expressed with 941 bp and 767 bp (that lacks 174 bp) which were translated into 32 kDa and 24 kDa precursor proteins, respectively. The 32 kDa precursor protein is processed to the 17 kDa mature IL-1alpha. Identical transcripts are also shown to be present in cat, dog and pig. Most of the functional role is assigned to the mature 17 kDa IL-1alpha isoform. However, functional analysis of recombinant rat IL-1alpha isoforms showed that there was a clear biopotency difference between these forms in order of 17 kDa IL-1alpha\u3e32proIL-1alpha\u3e24proIL-1alpha. Furthermore, the mature 17 kDa tIL-1alpha has also been implicated in pathologies such as orchitis, relapse of acute lymphoblastic leukemia (ALL) in the testis and infertility disorders in men. Thus, tIL-1alpha may play an important functional role both in coordination of normal testicular physiology as well as in contributing to the disease states in the testis
Impact of uteroplacental insufficiency on ovarian follicular pool in the rat
No full textAbstract Background A low oxygen supply to the fetus causes intrauterine growth restriction and can affect gonadal development of the offspring, having a potential impact on fertility. We investigated histology and gene expression in the postnatal rat ovary after fetal hypoxia induced by uterine artery ligation. Methods Sprague-Dawley rats underwent uterine artery ligation at day 19 of gestation. Offspring were sacrificed at 5, 20 and 40 days post-partum. Follicles were counted and classified in hematoxylin-eosin stained sections. Gene expression of 90 genes was analyzed by TaqMan® Low Density Array. Results A significantly lower number of total and primordial follicles was detected in 20 days post-partum intrauterine growth restricted animals. Follicle density was not different at 40 days post-partum, suggesting that compensatory mechanisms occurred during the pre-pubertal window. Uterine artery ligation modified the expression of 24 genes involved in different cellular functions, among which proliferation, apoptosis and metabolism. Conclusion Ovarian follicle pool was affected by fetal hypoxia in early life, but this effect did not persist in puberty. Genes involved in cellular processes were affected at all ages, potentially implying long-term genetic alterations. Further analyses are needed to elucidate later effects of fetal hypoxia on ovarian function and fertility
MeTeaM-A method for characterizing mature software metrics teams
No full textBackground: Metrics teams play an increasingly important role in handling data and information in modern software development organizations; they manage their companies' measurement programs, collect and process data, and develop and distribute information products. Metrics teams can comprise several roles, and their set-up can differ between companies, as can the metrics maturity of host organizations. These differences impact the effectiveness and quality of a team's measurement program. Objective: Our objective was to design and evaluate a model to describe the characteristics of a mature metrics team, which efficiently designs, develops, maintains, and evolves its organization's measurement program. Method: We conducted an action research study on four metrics teams of four distinct companies. We designed and evaluated a domain-specific model for assessing the maturity of metrics teams - MeTeaM - and also assessed the four metrics teams per se. Results: Our results were two-fold: the creation of the metrics team maturity model MeTeaM and a template to assess metrics teams. Our evaluation showed that the model captures the characteristics of successful metrics teams and quantifies the maturity status of both the metrics teams and their host organizations. Conclusions: More mature metrics teams score higher in the MeTeaM model than less mature teams. The assessment provides less mature metrics teams with valuable insights on what factors to improve. Such insights can be shared with and acted upon successfully with their organizations. (C) 2021 The Author(s). Published by Elsevier Inc
Mouse leydig cells with different androgen production potential are resistant to estrogenic stimuli but responsive to bisphenol a which attenuates testosterone metabolism.
No full textIt is well known that estrogens and estrogen-like endocrine disruptors can suppress steroidogenic gene expression, attenuate androgen production and decrease differentiation of adult Leydig cell lineage. However, there is no information about the possible link between the potency of Leydig cells to produce androgens and their sensitivity to estrogenic stimuli. Thus, the present study explored the relationship between androgen production potential of Leydig cells and their responsiveness to estrogenic compounds. To investigate this relationship we selected mouse genotypes contrasting in sex hormone levels and differing in testosterone/estradiol (T/E2) ratio. We found that two mouse genotypes, CBA/Lac and C57BL/6j have the highest and the lowest serum T/E2 ratio associated with increased serum LH level in C57BL/6j compared to CBA/Lac. Analysis of steroidogenic gene expression demonstrated significant upregulation of Cyp19 gene expression but coordinated suppression of LHR, StAR, 3βHSDI and Cyp17a1 in Leydig cells from C57BL/6j that was associated with attenuated androgen production in basal and hCG-stimulated conditions compared to CBA/Lac mice. These genotype-dependent differences in steroidogenesis were not linked to changes in the expression of estrogen receptors ERα and Gpr30, while ERβ expression was attenuated in Leydig cells from C57BL/6j compared to CBA/Lac. No effects of estrogenic agonists on steroidogenesis in Leydig cells from both genotypes were found. In contrast, xenoestrogen bisphenol A significantly potentiated hCG-activated androgen production by Leydig cells from C57BL/6j and CBA/Lac mice by suppressing conversion of testosterone into corresponding metabolite 5α-androstane-3α,17β-diol. All together our data indicate that developing mouse Leydig cells with different androgen production potential are resistant to estrogenic stimuli, while xenoestrogen BPA facilitates hCG-induced steroidogenesis in mouse Leydig cells via attenuation of testosterone metabolism. This cellular event can cause premature maturation of Leydig cells that may create abnormal intratesticular paracrine milieu and disturb proper development of germ cells
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